Microbiology Laboratory Manual

Adel M. Mahasneh, Salwa M. Bdour

Description

This manual is designed to satisfy the needs of students enrolled in؟ B.Sc. degree program in Biological, Microbiological, Agricultural and health professions. It provides؟ well balanced and chosen collection of relevant practical Microbiology Laboratory experiments. Students will perform experiments and report on quantitative as well as descriptive data pertaining to the concept they are tackling. The experiments in this manual stresses the quantitative methods, experimental controls, data analysis as well as report writing. The experiments were designed to provide maximum flexibility although each experiment represents؟ well defined concept, several experiments may be performed concurrently depending upon availability of tools and equipments as well as time constraints and students numbers in each laboratory session. Several appendixes appear at the end of the manual which include staining techniques, media composition and some bacterial diagnostic plates.

Sujets

• Sciences physiques
• Physique
• Sciences et techniques
• BIOLOGY

Informations

• Publié par: Academic for Publishing & Distribution Co.
• Date de parution: 01 janvier 2006
• Nombre de lectures: 11
• EAN13: 9796500145655
• Langue: English
• Poids de l'ouvrage: 2 Mo

Extrait

MICROBIOLOGY LABORATORY MANUAL

All right reserved Second Edition 2006 All right reserved no part of this book may be reproduced of transmitted in any means electronic or mechanical including system without the prior permission in writing of the publisher

Academics For Publishing & Distributing Co The Hashemite Kingdom of Jordan Amman Opposite of University Main Gate

Tel-fax: [phone] E-mail: [e-mail]

ISBN 9957-449-06-0

542.1 MAHASNEH, ADEL M. Microbiology Laboratory Manual / Adel Mahasneh, Salwa M. Bdour – nd 2 ed.- Amman -Academics For Publishing & Distributing, 2006 (124)P Size: 21 X 28 Deposit No.: 272/2/2006 Descriptors: / Microbiology// Laboratories//Higher Education/ Jordan// Manual/ Department of Press & Publications No. 286/2/2006

MICROBIOLOGY LABORATORY MANUAL

ADEL M. MAHASNEH Prof. of Microbiology

SALWA M. BDOUR Asso.Prof. Virology

UNIVERSITY OF JORDAN AMMAN

SECOND EDITION AMMAN 2006

MICROBIOLOGY- LABORATORY MANUAL

Preface This manual is designed to satisfy the needs of students enrolled in a B.Sc. degree program in Biological, Microbiological, Agricultural and health professions. It provides a well balanced and chosen collection of relevant practical Microbiology Laboratory experiments. Students will perform experiments and report on quantitative as well as descriptive data pertaining to the concept they are tackling. The experiments in this manual stresses the quantitative methods, experimental controls, data analysis as well as report writing. The experiments were designed to provide maximum flexibility although each experiment represents a well defined concept, several experiments may be performed concurrently depending upon availability of tools and equipments as well as time constraints and students numbers in each laboratory session. Several appendixes appear at the end of the manual which include staining techniques, media composition and some bacterial diagnostic plates. The authors welcome any useful comments and suggestions that may improve experiments and enhance their resolution. We also invite peers to recommend new experiments to be included in future editions if they see its relevance.

ADEL M. MAHASNEH SALWA M. BDOUR

5

6

MICROBIOLOGY- LABORATORY MANUAL

Contents

Subject Laboratory rules Microscopic observations of microorganisms in the living state Microscopic observation of simple stained bacteria Differential staining: The Gram stain Spore stain (Schaeffer- Fulton method) Capsule stain Total count a) Counting chamber method b) Breed’s method c) Indirect determination of total count by spectrophotometer Viable counts a) Preparation of serial dilution b) Pour plate method c) Spread plate method d) Miles an Misra method e) Membrane filter method Isolation of pure culture The microflora of air Conditions affecting growth of bacteria a) Microbial nutrient requirment b) Effects of sucrose concentration on growth of microorganisms c) Determination of oxygen requirment

Page 9 14

15 18 20 21 23 23 26 27

28 28 29 31 33 34 37 40 42 43 45

46

MICROBIOLOGY- LABORATORY MANUAL d) pH and microbial growth 49 e) The effect of various temperatures on growth of 50 microorganisms The lethal effect of temperature 52 The effect of ultraviolet light on the growth of bacteria 54 Antibiotics and bacterial growth (Susceptibility testing) 56 Antibiotic-resistant mutants 60 Oligodynamic action of heavy metals on bacteria 62 Microbial physiology 63 Production of acid and gas from sugars 64 Oxidation-fermentation of carbohydrates 66 Starch hydrolysis 68 Casein hydrolysis 70 Urea hydrolysis 71 Microbial population counts in soil 72 Ammonification in soil 74 Nitrification in soil 75 Nitrogen-fixation in soil 77 Microscopic examination of fungi 80 Agar block method 81 Disc method 83 Bacteriological examination of water 88 Isolation of fluorescent Pseudomonads 93 Isolation of lactic acid bacteria 95 Microbiological examination of pepper 96 Detection of Bacteriophages 98 Appendix I: Media Ingredients 101 Appendix II: Stains and Reagents 106

7

8

MICROBIOLOGY- LABORATORY MANUAL Appendix III: Use of Spectronic 20 Appendix IV: Celluler and Colonial Morphology Appendix V: Behavior of Bacteria on Diagnostic Media. References

110 111 113 124

MICROBIOLOGY- LABORATORY MANUAL MICROBIOLOGY LABORATORY LABORATORY RULES

1.Laboratory Coats must be worn in the Laboratory. Jackets , coats, books and other accessories not actually in use must be left in your locker in t the corridor; ON NO ACCOUNT SHOULD LAB. COATS BE WORN IN THF REFECTORY. 2.are not permitted in the laboratory. Do not drink fromSmoking and eating laboratory glassware. Moisten labels with water, not with the tongue. 3.In the case of accidents such as cuts, spilling live cultures, etc. report immediately to the Lecturer or Demonstrator. 4.cultures in the pails supplied. Never discard cultures downPlace discarded the sink. Even if you think that the organisms are harmless. Never remove material from the pails. Never take cultures out of the laboratory. 5.Two jars of DISINFECTANT are provided on each bench, one for PIPETTES and the other for USED SLIDES (Wet films only; on these the organisms are still alive). 6.Non –infections waste material such as FIXED AND STAINED SLIDES, burnt matches, waste paper etc. should be put into the pedal bins. Waste must not be put in the troughs or sinks. 7.Use only the microscope allocated to you (see list on notioeboard). Clean the instrument after use, paying particular attention to removal of oil from the oil immersion lens. using lens tissue. The tissue should be moistened with one drop of xylene if the oil has hardend on the objective. The oil should be removed in stages by gently polishing the objective with frsh pieces of lens tissue. Check the objective for oil also. 8.You must provide the following: a)1 box of 100 (3 in x 1 in). microscope slides b)1 fine pointed pental marker (not water-soluble). The colour will be specified at the first practical. c)1 hard-back laboratory notebook (A4 size) which will be examined and assessed at regular intervals. The reports of experiments should be written up in the following form:

9